TACS® 2 TdT Fluorescein Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Fix cells in methanol for 10 min. Incubate with 2% proteinase K solution for 15 minutes at room temperature. Incubate with 50 μL of Labeling Reaction Mix at 37˚C for 1 hour in a humidity chamber.	Analyze by fluorescence microscopy with a 495 nm filter.

Protocol tips
Fix cells in methanol for 10 min. Incubate with 2% proteinase K solution for 15 minutes at room temperature. Incubate with 50 μL of Labeling Reaction Mix at 37˚C for 1 hour in a humidity chamber.
Downstream tips
Analyze by fluorescence microscopy with a 495 nm filter.

Publication protocol

For the detection of neuronal apoptosis, the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end-labeling (TUNEL) assay was performed in both Ishikawa and HeLa cells using a commercial kit (TACS® 2 TdT-Fluor Apoptosis Detection Kit) according to the manufacturer's instructions. Briefly, the cells were immersion hydrated, and the fixed and immobilized samples were rinsed with a PBS solution for 10 minutes. The samples were pre-incubated with a 2% proteinase K solution for 15 minutes at room temperature before being incubated in a TdT labeling buffer for 5 minutes and with a labeling reaction mix for 60 minutes in a humidity chamber. After the cells were washed with PBS (0.05%), they were incubated for 30 min in the dark with a Strep-Fluor solution. After an additional washing period, the tissues were covered with a DAPI mounting medium (Vector, #H-1200) and analyzed by fluorescence microscopy with a 495 nm filter.

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Manufacturer protocol

Download the product protocol from Trevigen for TACS® 2 TdT Fluorescein Kit below.

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